用于下一代核酸定量的数字CRISPR系统。
Digital CRISPR Systems for the Next Generation of Nucleic Acid Quantification.
作者信息
Politza Anthony J, Nouri Reza, Guan Weihua
机构信息
Department of Biomedical Engineering, Pennsylvania State University, University Park 16802, USA.
Department of Electrical Engineering, Pennsylvania State University, University Park 16802, USA.
出版信息
Trends Analyt Chem. 2023 Feb;159. doi: 10.1016/j.trac.2023.116917. Epub 2023 Jan 6.
Digital CRISPR (dCRISPR) assays are an emerging platform of molecular diagnostics. Digital platforms introduce absolute quantification and increased sensitivity to bulk CRISPR assays. With ultra-specific targeting, isothermal operation, and rapid detection, dCRISPR systems are well-prepared to lead the field of molecular diagnostics. Here we summarized the common Cas proteins used in CRISPR detection assays. The methods of digital detection and critical performance factors are examined. We formed three strategies to frame the landscape of dCRISPR systems: (1) amplification free, (2) in-partition amplification, and (3) two-stage amplification. We also compared the performance of all systems through the limit of detection (LOD), testing time, and figure of merit (FOM). This work summarizes the details of digital CRISPR platforms to guide future development. We envision that improvements to LOD and dynamic range will position dCRISPR as the leading platform for the next generation of molecular biosensing.
数字CRISPR（dCRISPR）检测是一种新兴的分子诊断平台。数字平台为批量CRISPR检测引入了绝对定量并提高了灵敏度。凭借超特异性靶向、等温操作和快速检测，dCRISPR系统已做好引领分子诊断领域的准备。在此，我们总结了CRISPR检测中常用的Cas蛋白。研究了数字检测方法和关键性能因素。我们形成了三种策略来构建dCRISPR系统的格局：（1）无扩增，（2）分区内扩增，以及（3）两阶段扩增。我们还通过检测限（LOD）、检测时间和品质因数（FOM）比较了所有系统的性能。这项工作总结了数字CRISPR平台的细节，以指导未来的发展。我们设想，检测限和动态范围的改进将使dCRISPR成为下一代分子生物传感的领先平台。
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